Characterisation and identification of chemical constituents in aqueous extract of Fomes officinalis Ames based on ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry.

INTRODUCTION: Fungal species are an attractive resource for physiologically functional food and drug precursor. Fomes officinalis Ames, a medicinal fungus, is traditionally used as a folk medicine in traditional Chinese medicine prescription for the therapy of cough and asthma. The water-soluble substances in Chinese herbal medicines are likely to play an important physiological function. However, information on probing and identifying chemical components of the aqueous extract of Fomes officinalis Ames (AFO) remains unknown. OBJECTIVE: This study was conducted to screen and characterise the chemical components of AFO. MATERIAL AND METHODS: An effective and sensitive ultrahigh-performance liquid chromatography tandem quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method with the Full MS/PIL/dd-MS2 acquisition approach was applied for the profiling of chemical components in AFO. An HSS T3 column was used for component separation, and a strategy of simultaneous targeted and untargeted multicomponent characterisation was implemented. Multiple identification approaches were used, including accurate molecular mass and elemental composition matching, literature and database searching, and fragmentation rules elucidation. RESULTS: A total of 115 components, including 20 amino acids and derivatives, six nucleobases, nine nucleosides, 75 dipeptides, two tripeptides, and three other components, were tentatively identified. Among them, the targeted exploring method screened six nucleobases and nine nucleosides including modified nucleosides. To our best knowledge, this is the first time a report has been done on the presence of the 115 compounds in AFO. CONCLUSION: Profiling and characterisation compounds of AFO enriched its material basis, which would lay the foundation for improving potential medicinal and nutritional values and effecting comprehensive quality control of Fomes officinalis Ames.

Medicinal Potential of the Insoluble Extracted Fibers Isolated from the Fomes fomentarius (Agaricomycetes) Fruiting Bodies: A Review.

The tough, hoof-shaped fruiting bodies of the tinder conk mushroom, Fomes fomentarius (L.) Fr. (Polyporaceae, Agaricomycetes), were traditionally used all over the world as tinder to start fire, for ritual purposes, to make artworks like clothing, frames, ornaments, and also to cure various human diseases (wounds, gastro-intestinal disorders, liver-related problems, inflammations, various cancers, etc.). The first wave of scientific interest in F. fomentarius in Europe dates back to the early 1970s with the discovery of the red-brown pigments of the F. fomentarius external layer. Since then, a number of research papers and reviews have mentioned the history of use, taxonomy, composition and medicinal properties of some F. fomentarius preparations, e.g., soluble extracts and their fractions, isolated cell walls, mycelia and compounds purified from the culture broth. The present review is focused on the composition and benefits of the water-insoluble cell walls obtained from the F. fomentarius fruiting bodies. Isolated cell walls of the tinder mushroom reveal a fibrous hollow structure with an average diameter of 3-5 µm and a wall thickness of 0.2-1.5 µm. Naturally, the fibers are composed of 25-38% glucans, with a majority of ß-glucans, around 30% polyphenols, 6% chitin and less than 2% hemicellulose. The percentage of the main structural compounds can vary either slightly or considerably, depending on the extraction conditions. According to in vitro, in vivo, ex vivo as well as clinical studies, F. fomentarius fibers can modulate the immune system, contribute to intestinal health, accelerate wound healing, absorb heavy metals, organic dyes and radionuclides, normalize kidney and liver function, and provide antibacterial, antiviral, antifungal, anxiolytic, anti-inflammatory and analgesic effects. Multiple action of the insoluble cell walls purified from the F. fomentarius fruiting bodies is particularly effective in the treatment of chronic, recurring, complicated multifactorial diseases. It is certainly worth exploring the medicinal potential and the practical application of these preparations further.

Pectin decomposition at the early stage of brown-rot decay by Fomitopsis palustris.

The sapwood of Japanese cedar (Cryptomeria japonica D. Don) was decayed by the brown-rot fungus Fomitopsis palustris under bright and dark conditions. Scanning electron microscopy revealed the presence of mycelia inside the wood even after 1 week from the start of fungal exposure. Moreover, holes were observed in the torus after fungal exposure. Ruthenium red staining revealed that the pectin in pits was largely absent for 3 weeks. These events occurred before the mass loss of wood samples was confirmed at the early stage. Moreover, FpPG28A was more highly expressed at the hyphal front on a pectin-containing medium under dark conditions compared with bright conditions. This up-regulation under dark conditions indicated that the pectin decomposition ability was promoted inside the wood where light could not reach. In conclusion, we suggest that the brown-rot fungus completed its hyphal expansion within the wood via pectin decomposition in pits before holocellulose decomposition.

The HbA1c and blood glucose response to selenium-rich polysaccharide from Fomes fomentarius loaded solid lipid nanoparticles as a potential antidiabetic agent in rats.

Fomes fomentarius is a medicinal fungus used in traditional Chinese medicine to treat various illnesses. Antidiabetic effects of F. fomentarius extracts have been reported recently. In this study, F. fomentarius extracellular polysaccharide (PS) was prepared, and then to enhance its antidiabetic effects, Na2SeO3 was added to the culture medium, and selenium-polysaccharide (PS-Se) was obtained. Also, solid lipid nanoparticles containing PS (SLN-PS) and PS-Se (SLN-PS-Se) were synthesized by the microemulsion method to compare their effects with free polysaccharides in streptozotocin (STZ) diabetic rats. Optimized SLNs had a size of 170.5 nm and drug loading of 9.27 %. EDS analysis confirmed that Se presence in PS-Se. Characterization analyses such as FTIR, DSC, TGA, and XRD suggested that SLNs have good thermal stability and crystalline nature. Release of PS from SLNs demonstrated sustained profile, and MTT assay proved that PSs and SLNs have no cytotoxicity. Furthermore, oral administration of PS, PS-Se, SLN-PS, and SLN-PS-Se for 28 days to diabetic rats significantly declined blood glucose by 48.24 %, 49.96 %, 55.50 %, and 60.47 %, respectively. Also, insulin secretion and body weight improved, and HbA1c levels decreased. Treatment by PS, PS-Se, SLN-PS, and SLN-PS-Se alleviated lipid profiles, liver enzymes, and serum proteins. Liver anti-oxidant parameters and histopathological observation of the liver, pancreas, and kidney confirmed that F. fomentarius PSs and SLNs have antidiabetic impacts. Moreover, supplementation of PS with selenium improves its anti-hyperglycemic effects. Finally, SLN-PS and SLN-PS-Se showed a higher antidiabetic impact than free PS and PS-Se.

In Vitro Cytotoxic Activity of an Aqueous Alkali Extract of the Tinder Conk Mushroom, Fomes fomentarius (Agaricomycetes), on Murine Fibroblasts, Human Colorectal Adenocarcinoma, and Cutaneous Melanoma Cells.

Bioactive complexes of medicinal mushrooms have become attractive as complementary anticancer remedies. Our in vitro study focused on the cytotoxicity of the polyphenol-reach and beta-glucan-containing aqueous alkali extract from Fomes fomentarius fruiting bodies (FFE) using murine fibroblasts (L929), human colon adenocarcinoma cells (Caco-2), and cutaneous melanoma cells (COLO-818). Dose-dependent FFE cytotoxicity with an half maximal inhibitory concentration of 0.44 mg/mL was observed for L929 cells upon analysis of the total number of adherent cells, degree of cell viability, cell morphology, and mitochondrial metabolic activity. Cytotoxic effects on cancer cells tested using cell impedance were dependent on FFE concentration, type of cells, and their density. As a routine in vitro model for predicting human intestinal absorption, Caco-2 cells did not react on FFE, which can indirectly support its safety for the human intestinal epithelium. Melanoma cells were affected in a dose-dependent manner, even at low FFE concentrations (0.01-0.05 mg/mL). The confluent cell layer, which resembles a fully formed tumor, was much more resistant than the incompletely formed, subconfluent cell layer, simulating tumor formation. FFE applied topically could be a promising candidate to prevent melanoma development in its early stages.

Anti-oxidant and Anticancerous Effect of Fomitopsis officinalis (Vill. ex Fr. Bond. et Sing) Mushroom on Hepatocellular Carcinoma Cells In Vitro through NF-kB Pathway.

BACKGROUND: Fomitopsis officinalis (Vill. ex Fr. Bond. et Sing) is a medicinal mushroom, commonly called 'Agarikon'; it has traditionally been used to treat cough and asthma in the Mongolian population. OBJECTIVE: The objective of the study was to examine the significance of biological activity of F. officinalis and evaluation of the antioxidant activity and anticancer activity of six fractions of F. officinalis residues (Fo1-powder form dissolved in ethanol, Fo2-petroleum ether residue, Fo3-chloroformic, Fo4-ethylacetate, Fo5-buthanolic, and Fo6-waterethanolic) against hepatocellular carcinoma cells. METHODS: We performed in vitro studies of cell proliferation and viability assay, annexin V-FITC/Propidium Iodide assay, and NF-kB signaling pathway by immunoblot analysis. RESULTS: Our findings revealed that all six fractions/extracts have antioxidant activity, and somehow, they exert anticancerous effects against cancer cells. In cancerous cell lines (HepG2 and LO2), Fo3 chloroformic extract promoted the cancer cell apoptosis and cell viability, activated G2/M-phase cell cycle, and selectively induced NF-kB proteins, revealing as a novel antitumor extract. CONCLUSION: This study reports that Fo3-chloroformic extract is rich in antitumor activity, which was previously not investigated in cancer. To develop the impact of F. officinalis among natural products to treat/prevent oxidative stress disorders or cancers, further examinations of F. officinalis are needed to develop new natural drugs to treat cancer. However, this study assessed only one extract, Fo3-chloroformic, which has a significant impact against cancer cell lines.

Predicting multi-enzyme inhibition in the arachidonic acid metabolic network by Heritiera fomes extracts.

Heritiera fomes is a mangrove plant with a rich history of ethnomedicinal usage against chronic inflammation. Biochemical analyses of H. fomes have exposed a plethora of bioactive phytochemicals that contribute to this therapeutic effect by perturbing enzymes of a complex inflammatory network mediated by arachidonic acid (AA) metabolism. This study is the first instance of utilizing cheminformatic approaches to elucidate a molecular linkage between these phytochemical interventions and the multi-enzyme AA metabolic network regulation. Analysis of the simulations reflects H. fomes as a functional reservoir of multiple safe and potent natural anti-inflammatory compounds. The investigation suggests two phytocompounds extracted from the plant: a sesquiterpene lactone and a flavone glycoside, as candidate inhibitors of multiple catalytic checkpoints of the inflammatory network. The outcomes of this research act as a primary guideline for future laboratory and clinical testing of anti-inflammatory potentials of H. fomes as an exploitable source of safe and potent drug-like molecules.Communicated by Ramaswamy H. Sarma.

Fortified Mycelium of Fomitopsis officinalis (Agaricomycetes) as a Source of Biologically Active Substances Effective in the Prevention of Civilization Diseases.

Fomitopsis officinalis is a popular species of therapeutic arboreal mushroom with a documented history of use in traditional European and Asian medicine. It was chosen as a research material not only due to its anti-inflammatory, antimicrobial, and immunostimulatory properties but also because it is endangered in Poland. The aim of this study was to determine the effects of supplementing inorganic zinc and magnesium salts to the culture medium in which F. officinalis was multiplied in vitro in 10-L bioreactors equipped with an aeration and CO2 removal system. The influence of the addition of zinc and magnesium salts to the culture medium on the quantity of bioelements and organic compounds (indole, phenolic compounds, and L-phenylalanine), as well as their bioavailability to the human body (extraction to artificial digestive juices), was determined. The lowest amount of magnesium and zinc was found in the fruiting bodies of F. officinalis (132.1 and 15.3 mg/100 g dry weight [DW], respectively), whereas the highest amount was found in the mycelium obtained on the medium enriched with sulfate salts (1261 and 182.8 mg/100 g DW, respectively). Enrichment of the medium increased the content of the studied bioelements and organic compounds in the F. officinalis mycelium. The results showed that the applied method allowed obtaining fortified mycelium of the tested species as a natural therapeutic material that can supplement the deficiency of bioelements, phenolic compounds, L-phenylalanine, and indole compounds.

Effect of Cytokinin-Containing Extracts from Some Medicinal Mushroom Mycelia on HepG2 Cells In Vitro.

Endogenous cytokinins in mycelia of medicinal mushrooms Hericium coralloides and Fomitopsis officinalis grown in vitro were identified using high-performance liquid chromatography coupled with mass spectrometry. High amounts of zeatin-type cytokinins and isopentenyladenine were found. The qualitative composition and quantitative content of cytokinins were species-specific traits of mushrooms. Optical microscopy was used to perform a comparison analysis of the influence of crude extracts and purified cytokinin fractions from both species' mycelial biomass on HepG2 tumor cell growth in vitro and morphology. The results showed that purified cytokinin fractions from H. coralloides and F. officinalis mycelia demonstrated a cytotoxic effect on HepG2 cells, unlike crude extracts. Under the influence of all mushroom extracts, similar patterns of changes in HepG2 cell morphology were observed, but they were more pronounced for H. coralloides compared with F. officinalis. Purified fractions of both mushroom species caused an increased level of apoptosis compared to crude extracts. Some increase in glucose uptake by cultured cells was found in all investigated samples wherein the influence of H. coralloides extracts was approximately twice the effect of the corresponding F. officinalis extracts. The data obtained confirm the assumption that cytokinins are involved in the expression of therapeutic effects of medicinal mushrooms and indicate the need to take into consideration the methods of cytokinin extraction when preparing pharmacologically active drugs based on fungal raw materials. Thus, extracts from H. coralloides and F. officinalis mycelial biomass are promising in the search for anticancer agents.

Chemical Composition and Ultraviolet Absorption Activity of an Aqueous Alkali Extract from the Fruiting Bodies of the Tinder Conk Mushroom, Fomes fomentarius (Agaricomycetes).

White rot mushroom Fomes fomentarius is a medicinal fungus with great potential to be explored. This work focused on the chemical composition of a basic aqueous extract from F. fomentarius fruiting bodies. The extract was mostly composed of phenolics, carbohydrates, minerals, and crude fat with a low amount of proteins and chitin. One-third of the total carbohydrates were in the form of beta-glucans with minor amounts of alpha-glucans. The most valuable essential part of the extract was composed of an acid-resistant ultraviolet (UV)-absorbing mixture of phenolic compounds such as melanins, lignins, and humic acids. These compounds, also referred to as melanin-like pigments, provided for the high antioxidant activity of the extract measured in vitro. Moderate sun-protective capacity was observed with regard to UVB rays and also expected in the UVA range. Quantification of melanin-like pigments in the F. fomentarius extract was possible either gravimetrically as acid-insoluble residue or spectrophotometrically in the UV region. Melanin estimation, based on nitrogen measurements, offered misleading results due to the presence of nitrogen-free melanins along with other nitrogen-containing compounds such as proteins and chitin. F. fomentarius water-soluble basic extract, containing beta-glucans and rich in melanin-like substances, could be used, for example, for topical skin application to prevent cell damage caused by excessive UV exposure or cytotoxic free radicals. The bioactive potential, safety, and further applications of the F. fomentarius extract are currently being investigated.

Simplified Colorimetric Determination of Glucosamine in the Tinder Conk Mushroom, Fomes fomentarius (Agaricomycetes).

Next to cellulose, chitin is the most abundant biopolymer on Earth and participates, in particular, in the assembly of the fungal cell walls. Easy monitoring of the amount of chitinous polymers could be advantageous during biotechnological cultivation and mushroom farming in order to control and optimize the production process. In this study, a modified colorimetric glucosamine (GlcN) assay was validated using high-performance anion exchange chromatography (HPAE) coupled with pulsed amperometric detection (PAD) following acid hydrolysis of the Fomes fomentarius cell walls. The average data obtained from both methods were well comparable. The HPAE-PAD analysis was suggested to be preferable because it had less data scattering, but it could be replaced with the simplified colorimetric GlcN screening in some cases such as low budgetary resources or an absence of suitable equipment, materials, or highly skilled staff. The robust, cost-effective colorimetric GlcN assay presented here can be readily requested, considering the rapidly growing market as well as the scientific and industrial potential of gourmet and medicinal mushrooms.

Decolorization and biodegradation of melanoidin contained in beet molasses by an anamorphic strain of Bjerkandera adusta CCBAS930 and its mutants.

We used a ligninolytic strain of the white-rot fungus B. adusta CCBAS 930 and its mutants with modified ligninolytic activity to assess their potential to remove of molasses. The analyzed strains have been shown to be able to decolorize 1% or 2% molasses solutions containing brown-colored toxic melanoidins. It was found that the decolorization process was determined by the transition to the stage of production of sporulating aerial mycelium (liquid and agar cultures) coupled with an increase in peroxidase activity, which was accompanied by a decrease in the level of melanoidin, free radicals, and phenolic compounds. Four different peroxidase activities were detected in post-culture liquids, i.e. horseradish-like (HRP-like), manganese-dependent (MnP), lignin (LiP), and versatile (VP) peroxidase activities. The HRP-like peroxidase was characterized by the highest activity. The efficiency of removal of melanoidins from a 1% molasses solution by the parental strain and the mutants was dependent on the culture method. The highest efficiency was noted in immobilized cultures (threefold higher than in the mycelium-free cultures), which was accompanied by stimulation of HRP-like peroxidase activity. Mutant 930-5 was found to be the most effective in the decolorization and decomposition of melanoidin. The HRP-like activity in the immobilized cultures of B. adusta 930-5 was 640-fold higher than in the mycelium-free cultures of the fungus. Moreover, decolorization and biodegradation of melanoidin by B. adusta CCBAS 930 and 930-5 was coupled with detoxification.

Medicinal potential of mycelium and fruiting bodies of an arboreal mushroom Fomitopsis officinalis in therapy of lifestyle diseases.

Fomitopsis officinalis is a medicinal mushroom used in traditional European eighteenth and nineteenth century folk medicine. Fruiting bodies of F. officinalis were collected from the natural environment of Swietokrzyskie Province with the consent of the General Director for Environmental Protection in Warsaw. Mycelial cultures were obtained from fragments of F. officinalis fruiting bodies. The taxonomic position of the mushroom mycelium was confirmed using the PCR method. The presence of organic compounds was determined by HPLC-DAD analysis. Bioelements were determined by AF-AAS. The biochemical composition of the tested mushroom material was confirmed with the FTIR method. Antioxidant properties were determined using the DPPH method, and the antiproliferative activity was assessed with the use of the MTT test. The presence of indole compounds (L-tryptophan, 6-methyl-D,L-tryptophan, melatonin, 5-hydroxy-L-tryptophan), phenolic compounds (p-hydroxybenzoic acid, gallic acid, catechin, phenylalanine), and sterols (ergosterol, ergosterol peroxide) as well as trace elements was confirmed in the mycelium and fruiting bodies of F. officinalis. Importantly, a high level of 5-hydroxy-L-tryptophan in in vitro mycelium cultures (517.99 mg/100 g d.w) was recorded for the first time. The tested mushroom extracts also showed antioxidant and antiproliferative effects on the A549 lung cancer cell line, the DU145 prostate cancer cell line, and the A375 melanoma cell line.

Lanostane Triterpenoids with PTP1B Inhibitory and Glucose-Uptake Stimulatory Activities from Mushroom Fomitopsis pinicola Collected in North America.

A chemical investigation on the fruiting bodies of Fomitopsis pinicola led to the isolation and identification of 28 lanostane triterpenoids including 11 new compounds (1-11) and 17 known analogues (12-28). Their structures were elucidated by extensive one-dimensional NMR, two-dimensional NMR, and MS spectra. All isolates were tested for their anti-inflammatory activity, protein tyrosine phosphatase 1B (PTP1B) inhibitory activity in vitro, and effect on glucose uptake in insulin-resistant HepG2 cells. Compounds 1, 4, 22, 23, and 27 inhibited the nitric oxide released from the LPS-induced RAW 264.7 cell assay with IC50 values in the range of 21.4-27.2 µM. Compounds 18, 22, 23, and 28 showed strong PTP1B inhibitory activity with IC50 values in the range of 20.5-29.9 µM, comparable to that of the positive control of oleanolic acid (15.0 µM). Compounds 18 and 22 were confirmed to be good competitive inhibitors of PTP1B by kinetic analysis. In addition, compounds 18, 22, and 28 were found to stimulate glucose uptake in the insulin-resistant HepG2 cells in the dose from 6.25 to 100 µM. These findings indicated the potential of F. pinicola in the development of functional food or medicine for the prevention and treatment of diabetes.

Fomitopsis officinalis: a Species of Arboreal Mushroom with Promising Biological and Medicinal Properties.

Medicinal mushrooms of the order Polyporales have a long history of use, which is evidenced by the finding of dissected fruiting bodies with Ötzi, who lived over 5000 years ago. Because of its valuable biological properties and its use in 18th and 19th-century pharmacy, Fomitopsis officinalis used to be mass-collected. Moreover, the large demand for larch wood and non-wood materials (resin) caused an excessive exploitation of larch forests, which directly contributed to the disappearance of F. officinalis from its natural environment. The qualities of medicinal preparations obtained from the F. officinalis fruiting bodies are determined by the unique composition of its bioactive compounds, such as: triterpenoids, polysaccharides, organic acids, coumarins and phenolic compounds. It has been proved that both crude extracts and the compounds isolated from F. officinalis have a wide spectrum of therapeutic effects, including anti-inflammatory, cytotoxic, and antimicrobial effects.

Characterisation of Extracts and Anti-Cancer Activities of Fomitopsis pinicola.

Fomitopsis pinicola (Sw. Karst) is a common bracket fungus, with a woody texture. It is found predominantly in coniferous forests in temperate regions throughout Europe and Asia. Fomitopsis pinicola has been extensively used for medicinal purposes, particularly in Chinese and Korean traditional medicine. In this mini-review, the anti-cancer characteristics of F. pinicola extracts were investigated. In vitro experiments revealed the pro-apoptotic, anti-oxidant and anti-inflammatory properties of extracts, whilst two of three in vivo studies reported an inhibition of tumour growth and prolonged survival. Only studies wherein fungal specimens were sourced from Europe or Asia were included in this review, as samples sourced as F. pinicola from North America were probably not F. pinicola, but a different species. Although not one of the most revered fungal species, F. pinicola has been used as a medicinal fungus for centuries, as well as consumed as a health food supplement. To date, the results from only three in vivo studies, investigating anti-cancer properties, have been published. Further studies, using comprehensively identified specimens, are required to fully elucidate the anti-cancer properties of F. pinicola extracts.

Antidiabetic Properties of the Red Belt Conk Medicinal Mushroom Fomitopsis pinicola (Agaricomycetes) Extracts on Streptozotocin-Induced Diabetic Rats.

The antidiabetic effect of different doses of water extract (WE) and ethanol extract (EE) was tested on a high-fat diet and streptozotocin (STZ) induced diabetic rats. Parameters were evaluated with normal control (NC), diabetes mellitus control (DM), and metformin (M) groups. In the experiment, nine groups were used with eight rats in each group and three doses of each WE and EE were used, with low, medium, and high doses. The results revealed that the DM group lost a significant amount of weight, whereas the NC group's weight increased throughout the experiment. After treatment with Fomitopsis pinicola, the EE group's weight increased gradually. Liver, kidney, and pancreas weight decreased after STZ injection and returned to normal in EE treated groups. Fasting blood glucose (FBG) levels were observed to be significantly lower after F. pinicola treatment. Serum insulin levels were also restored to normal after mushroom extracts supplementation. Specifically, STZ-induced hyperglycemia was inhibited by high dose EE administration. The biochemical analysis revealed that high-dose EE treatment increased HDL-C and decreased TC, TG, and LDL-C. Results demonstrated that high-dose EE administration protected the organ tissues from oxidative stress by normalizing the antioxidant levels, and CAT, SOD, and GSH-Px suppressed the lethal effect of MDA. The study concluded that F. pinicola EE at the dose 300 mg/kg has a more hypoglycemic, hyperinsulinemic, antioxidant, and antihyperlipidemic effect than NC, DM, and M, and regulates hyperglycemia by increasing insulin secretion.

Cultivation and Medicinal Value of the Red Belt Conk Mushroom Fomitopsis pinicola (Agaricomycetes).

This study examined biological characteristics, liquid fermentation, and cultivation of Fomitopsis pinicola. A single-factor test concluded that the optimal carbon and nitrogen sources for mycelial growth were soluble starch and yeast paste; the optimal culture temperature was 31°C, and the optimal pH was 6.0. The orthogonal experiment indicated that the optimal formula for mycelial culture was 25 g soluble starch, 2 g yeast extract, 1 g KH2PO4, and 1.5 g MgSO4 added to 1 L water. The optimal conditions for liquid fermentation culture consisted of the following: a loading volume 90 mL, inoculation volume 30 mL, and rotation speed 160 rpm. The optimal substrate formula for domestic culture was 20% corn cob, 30% sawdust, 20% wheat bran, 25% cotton seed shell, 3% corn meal, 1% gypsum, and 1% lime, which produced the highest yield of fruiting bodies. The results provided basic data for deep liquid fermentation culture and recommendations for the further development and utilization of F. pinicola.

Anticancer Characteristics of Fomitopsis pinicola Extract in a Xenograft Mouse Model-a Preliminary Study.

Introduction: Medicinal mushrooms have been used for the treatment of diseases and general promotion of health for many centuries. Recent pharmacological research into medicinal mushrooms has identified various therapeutic properties, with applications in modern medicine.Aim: To evaluate the anti-cancer activities of Fomitopsis pinicola (F. pinicola) alcoholic extract in an in vivo setting.Methods: The anti-tumour effect of the F. pinicola extract was tested in a xenograft immune-compromised Rag-1 mouse model. This was followed by RT-PCR and metabolomics analyses.Results: There were no observable differences in tumor growth between treated and non-treated groups. The bioactive components were not detected in the mouse plasma or the tumor site.Conclusions: The extract was poorly absorbed; this is likely due to the timing of treatment, dosage levels and modifications made to the extract where the alcohol-based solvent was replaced with water. This, in combination with fractionation studies which identified most anti-cancer compounds to be hydrophobic, largely explained the lack of anti-cancer activities in vivo.

C30 and C31 Triterpenoids and Triterpene Sugar Esters with Cytotoxic Activities from Edible Mushroom Fomitopsis pinicola (Sw. Ex Fr.) Krast.

Fomitopsis pinicola (Sw. Ex Fr.) Krast has been commonly used as a health food source and antitumor agent. To uncover bioactive key composition of F. pinicola, in our study, we investigated the chemical constituents of a methanol extract of F. pinicola and thirty-five lanostane-type tritetpenoids; 13 new compounds (1-13) and twenty-two known analogues (14-35) were isolated. Among them, compounds 1-9 were C30 lanostane triterpenoids and triterpene sugar esters, while compounds 10-13 were C31 triterpenoids and triterpene sugar esters. Their structures and absolute configurations were elucidated by extensive 1D, 2D NMR, MS, and IR spectra. Furthermore, cytotoxic activities of all isolates against five human tumor cell lines (HL-60, A549, SMMC-7721, MCF-7, and SW480) were evaluated. The results showed that compounds 12, 14, 17, 18, 22, and 23 displayed cytotoxic effects against five human tumor cell lines with IC50 values ranging from 3.92-28.51 µM. Meanwhile, compounds 9 and 35 exhibited selected inhibitory activities against HL-60, SMMC-7721, and MCF-7 with IC50 values in the range of 13.57-36.01 µM. Furthermore, the flow cytometry analysis revealed that compounds 17, 22, and 35 induced apoptosis in HL-60 cell lines. Their structure-activity relationships were preliminarily reported. These findings indicate the vital role of triterpenoids and their glycosides in explaining antitumor effects of F. pinicola and provide important evidence for further development and utilization of this fungus.